Transcriptome analysis of Arabidopsis thaliana WT, mbd1 K.O., pyl5 K.O. and mbd1 pyl5 DOUBLE K.O. mutants

Transcriptome profiling of WT, mbd1, pyl5 and mbd1 pyl5 knock-out mutants in response to Abscisic acid (ABA) was done using Affymetrix GenechipTM Arabidopsis ATH1 Genome Array. A comparative analysis of the transcriptome profiles of WT, mbd1, pyl5 and mbd1 pyl5 in response to ABA was performed. To begin with, WT, mbd1, pyl5 and mbd1 pyl5 plants were grown in ½ MS medium either lacking or supplemented with 0.3 M ABA. Total RNA was extracted from one-week old seedlings and the quality and quantity of the RNA was determined by Agilent Bioanalyzer and Nanodrop spectrophotometer. Three biological replicates per sample were used for WT and mbd1 (i.e. three for ½ MS medium and three for ABA-containing medium) while two biological replicates per sample were used for pyl5 and mbd1 pyl5. Preparation of labelled cRNA and array hybridization was carried out as per manufacturer’s instructions.